Antiviral composition comprising ABCG transporter activator and use thereof

Abstract

The present invention relates to: an antiviral composition against coronavirus, comprising an ABCG transporter activator; and use thereof. The composition comprising a ginseng seed oil-in-water nanoemulsion, of the present invention, has been confirmed to have a significant antiviral effect against coronavirus by reactivating the expression of ABCG transporters suppressed by coronavirus. Therefore, the composition can be effectively used in the fields of pharmaceuticals, quasi-drugs, foods, cosmetics, and the like, for treating, preventing, and ameliorating coronavirus infectious diseases or treating, preventing, and ameliorating inflammatory diseases caused by coronavirus.

Description

Antiviral composition comprising ABCG transporter activator and use thereof

[0001] The present invention relates to an antiviral composition against a coronavirus comprising an ABCG transporter activator and its use.

[0002] The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which causes COVID-19, is characterized by the development of potentially fatal pneumonia. Symptoms such as cough, cold-like symptoms, shortness of breath, chills, and sore throat can appear within 2-14 days of exposure. The disease rapidly progresses to systemic hypoxia and acute respiratory syndrome. Despite recent declines in the spread of the coronavirus, complete eradication remains a possibility, and the risk of a large-scale epidemic remains high, necessitating the development of therapeutics. Understanding the biological regulation of COVID-19 within host cells is therefore a crucial task. Furthermore, there is a critical need for personalized therapies and diverse drug delivery methods to promote recovery from infection while minimizing side effects.

[0003] Generally, Biosafety Levels (BSLs) are classified based on the potential hazard posed by a specific pathogen, ranging from BSL-1 (lowest risk) to BSL-4 (highest risk). Coronaviruses fall under BSL-3, which requires a higher level of isolation, strict safety measures, and specialized research facilities. Human coronavirus HCoV-OC43, which belongs to the same virus genus as SARS-CoV and SARS-CoV-2, is a human pathogen distinct from SARS-CoV-2. Human coronaviruses are known to cause the common cold. Human coronavirus HCoV-OC43 is the most common human coronavirus and is known to cause upper respiratory tract infections. Recent studies have shown that a large number of children seroconvert to human coronaviruses early in life before the age of two, with Human coronavirus HCoV-OC43 being the most common. Human coronavirus (HCoV)-OC43 infection has been implicated in the development of severe respiratory disease in children and the elderly, and has been confirmed to cause severe lower respiratory disease in children (Pediatr. Infect. Dis. J., 32(4):325-329). Because no specific antiviral treatment exists for HCoV-OC43, the need for antiviral agents targeting HCoV-OC43 and related coronaviruses to alleviate disease and prevent severe outcomes is emphasized.

[0004]

[0005] Meanwhile, in the recent paradigm of the food industry, the deployment of functional food ingredients is crucial for enhancing the nutritional efficacy and preventative potential of dietary products. These ingredients include cholesterol regulation, glucose homeostasis, and antioxidant defense mechanisms. These ingredients not only enhance the inherent properties of foods but also align with the rapidly growing consumer preference for nutritionally enhanced and functional foods. These advancements aim to improve food safety, promote the acceptance of functional foods, ensure food safety through better traceability and risk management, and support sustainable food production practices.

[0006] Furthermore, in recent years, there have been significant advances in biomedical research focused on the application of advanced food-derived drug delivery systems. This growing demand is primarily driven by the growing need for improved therapeutic efficacy, minimal side effects, and improved patient compliance with treatment regimens. Among these innovative delivery systems, nanoemulsions are emerging as a flexible and promising platform for addressing various challenges faced in pharmaceutical formulation, food science, cosmetics, and other industries.

[0007]

[0008] Meanwhile, ginseng (Panax ginseng CA Meyer) is a perennial herbaceous plant of the Araliaceae family, primarily cultivated in northeastern China, South Korea, and eastern Russia. Ginseng has long been considered a valuable resource, serving as both a source of medicine and food. Modern medical research has revealed that ginseng possesses significant biological activities and therapeutic effects, including neuroprotective, anticancer, cardioprotective, and antioxidant properties. In recent years, ginseng seed oil (GSO) has attracted attention as a source of novel bioactive ingredients with diverse potential applications in health and wellness.

[0009] However, no research has yet been conducted to optimize the formulation of ginseng seed oil into a nanoemulsion. Further research is needed to determine its efficacy against various viruses, optimal concentration, delivery method, and interactions with different viruses. Furthermore, there is no research on target sites for superior antiviral activity against coronaviruses.

[0010] One object of the present invention is to provide an ABCG transporter activator comprising ginseng seed oil.

[0011] Another object of the present invention is to provide an antiviral composition against coronavirus comprising an ABCG transporter activator.

[0012] Another object of the present invention is to provide a pharmaceutical composition for preventing or treating a coronavirus infectious disease, comprising the antiviral composition.

[0013] Another object of the present invention is to provide a method for preventing or treating a coronavirus infectious disease, comprising administering the pharmaceutical composition to a subject.

[0014] Another object of the present invention is to provide a health functional food composition for preventing or improving coronavirus infectious disease, comprising the antiviral composition.

[0015] Another object of the present invention is to provide a pharmaceutical composition for preventing or improving a viral infection disease, comprising the antiviral composition.

[0016] Another object of the present invention is to provide a feed composition for preventing or improving a viral infection disease, comprising the antiviral composition.

[0017] Another object of the present invention is to provide a pharmaceutical composition for preventing or treating an inflammatory disease caused by a coronavirus, comprising the antiviral composition.

[0018] Another object of the present invention is to provide a method for preventing or treating an inflammatory disease caused by a coronavirus, comprising administering the pharmaceutical composition to a subject.

[0019] Another object of the present invention is to provide an anti-inflammatory composition comprising the antiviral composition.

[0020] Another object of the present invention is to provide a health functional food composition for preventing or improving inflammatory diseases, comprising the antiviral composition.

[0021] Another object of the present invention is to provide a pharmaceutical composition for preventing or improving an inflammatory disease caused by a coronavirus, comprising the antiviral composition.

[0022] Another object of the present invention is to provide a method for preparing an antiviral composition against coronavirus comprising an ABCG transporter activator comprising a ginseng seed oil oil-in-water nanoemulsion, the method comprising a step of formulating ginseng seed oil into an oil-in-water nanoemulsion.

[0023] Another object of the present invention is to provide a use for activating ABCG transporter comprising ginseng seed oil.

[0024] Another object of the present invention is to provide an antiviral use of a composition comprising an ABCG transporter activator against coronaviruses.

[0025] Another object of the present invention is to provide a composition comprising an ABCG transporter activator for use in preventing, improving or treating coronavirus infectious diseases.

[0026] Another object of the present invention is to provide an anti-inflammatory use of a composition comprising an ABCG transporter activator.

[0027] The composition comprising the ginseng seed oil-in-water nanoemulsion of the present invention was confirmed to have a remarkable antiviral effect against coronavirus by reactivating the expression of the ABCG transporter suppressed by coronavirus, and thus can be usefully utilized in the fields of pharmaceuticals, quasi-drugs, foods, and cosmetics for the treatment, prevention, and improvement of coronavirus infectious diseases or the treatment, prevention, and improvement of inflammatory diseases caused by coronavirus.

[0028] Figure 1 is a diagram confirming the optimal GSO concentration and surfactant ratio for confirming GSO-NE stability according to one embodiment of the present invention. Specifically, it shows the results of zeta potential analysis using GSO ratios (5%, 10%, 15%, and 20%) and surfactant ratios (1:1, 2:1, 3:1, and 4:1).

[0029] FIG. 2 is a diagram confirming the optimization of the GSO-NE formulation according to one embodiment of the present invention. Specifically, AB measured the Z-average, zeta potential, and polydispersity index (PDI) using different amplitude percentages (20%, 40%, 60%, and 80%) at various time intervals (5, 10, 20, and 40 minutes), and CD is a diagram confirming the durability of GSO-NE over 1 to 6 months at various storage temperatures (4°C, 25°C, and 37°C) by measuring the Z-average, zeta potential, and PDI. G is a diagram confirming the final optimal conditions of the GSO-NE of the present invention, in which the Z-average is (nm) 359.6, the PDI is 0.24, and the zeta potential (mV) is -37.01.

[0030] FIG. 3 is a diagram confirming the cytotoxicity of GSO-NE according to one embodiment of the present invention for HCoV-OC43 infection in Vero E6 cells and confirming its effect. Specifically, A is the result of cytotoxicity analysis of GSO-NE (0.625, 1.25, 2.5, 5, 10, 20, and 40 μg / mL) in Vero E6 cells, B shows the IC50 value of GSO-NE treated in Vero E6 cells, C shows the morphological image after treating GSO-NE (2.5 and 5 μg / mL) in Vero E6 cells infected with HCoV-OC43, and D is a diagram confirming HCoV-OC43 RNA expression through PCR in Vero E6 cells treated with various concentrations of GSO-NE (0.625, 1.25, 2.5, and 5 μg / mL). In addition, EG confirmed the expression of inflammatory cytokines (IL-6, TNF-α, IL-1β, and MCP-1) in the Mock (control) group, HCoV-OC43-infected group, and GSO-NE treatment (2.5 and 5 μg / mL) groups in Vero E6 cells (*p < 0.05, **p < 0.01, and ***p < 0.001, compared with the untreated Mock group).

[0031] Figure 4 shows the results of transcriptome analysis that identified differentially expressed genes in GSO-NE according to one embodiment of the present invention. Specifically, A represents genes with significantly increased and decreased expression (fold change value >±1.0) in a volcano plot, with gold indicating significance and black indicating insignificance. The bar graph indicates the number of increased and decreased genes. B represents the results of exploring functional pathways through KEGG analysis, and C represents the results of gene ontology analysis through classification of differentially expressed genes into biological processes (BP), cellular components (CC), and molecular functions (MF).

[0032] Figure 5 illustrates a functional analysis of the ATP-binding cassette (ABC) receptor gene family. Specifically, A is a heatmap showing the fold change values ​​of the ABCA, ABCB, ABCC, ABCD, and ABCG protein families between HCoV-OC43 and GSO-NE; B is a visualization of the protein-protein interaction network of the ABCA, ABCB, ABCC, ABCC, ABCD, and ABCG protein families; and C is a diagram showing the ABCG protein family using the STRING server.

[0033] Figure 6 is a diagram showing the regulation of ABCG genes of the ABC transporter subfamily treated with GSO-NE according to one embodiment of the present invention in Vero E6 cells. Specifically, A-E are bar graphs showing the mRNA expression levels of ABCG1, ABCG2, ABCG4, ABCG5, and ABCG8 detected through qRT-PCR analysis, and F is a Western blotting image showing the protein expression levels of ABCG1, ABCG5, and ABCG8 after treatment with GSO-NE. Here, each bar corresponds to the mean ± standard error obtained from three independent experiments using the same sample, and statistical significance is indicated by an asterisk (*p < 0.05, **p < 0.01, ***p < 0.001, compared with the untreated Mock group).

[0034] Hereinafter, the present invention will be described in more detail.

[0035]

[0036] Meanwhile, each description and embodiment disclosed in the present invention can also be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in the present invention fall within the scope of the present invention. Furthermore, the scope of the present invention should not be considered limited by the specific descriptions described below.

[0037] Furthermore, those skilled in the art will recognize or be able to ascertain, using no more than routine experimentation, numerous equivalents to the specific embodiments of the invention described herein. Furthermore, such equivalents are intended to be encompassed by the present invention.

[0038] Additionally, numerous papers and patents are referenced and cited throughout this specification. The disclosures of these cited papers and patents are incorporated herein by reference in their entirety to provide a clearer understanding of the state of the art and the scope of the present invention.

[0039]

[0040] To achieve the above purpose, one aspect of the present invention provides an ABCG transporter activator comprising ginseng seed oil.

[0041] Another aspect of the present invention provides a composition for activating an ABCG transporter comprising ginseng seed oil.

[0042]

[0043] In the present invention, “ABCG transporter” refers to the G subfamily of ATP-binding cassette (ABC) transporters.

[0044] In the present invention, "ABCG transporter activator" means an agent that activates the expression or activity of the ABCG transporter, and means activating, increasing expression, or reactivating expression suppressed by a coronavirus. Specifically, it includes all substances that can activate, increase, or promote the production and expression of an ABCG transporter protein or a polynucleotide encoding an ABCG transporter protein, or a transcript derived therefrom, or re-express, re-increase, re-promote, or reactivate expression or activity suppressed by a coronavirus. The activator may be used in combination with a stimulator or an enhancer, and the activating agent may include an expression enhancer, an activity enhancer, a reactivator that reactivates what has been suppressed by a coronavirus, etc. The novel ABCG transporter activator of the present invention may be used in combination with other known ABCG transporter activators.

[0045] As an example of implementation, the present inventors discovered a formulation optimized for the prevention or treatment of coronavirus infectious diseases, including ginseng seed oil as a novel ABCG transporter activator, and particularly ginseng seed oil in the form of an oil-in-water nanoemulsion.

[0046] In particular, the inventors of the present invention confirmed that the ginseng seed oil-in-water nanoemulsion effectively inhibits coronavirus by reversing, i.e., reactivating, the expression of the ABCG transporter inhibited by coronavirus, for the prevention or treatment of coronavirus infectious diseases.

[0047]

[0048] In the present invention, the term "ginseng seed oil" refers to oil extracted from the seeds of ginseng (Panax ginseng CAMeyer). Ginseng is a plant belonging to the Araliaceae family, and is a perennial plant that grows very slowly compared to other plants, and refers to a plant mostly found in Korea, the northeastern part of China, or the Siberian Far East. Specifically, the ginseng may be selected from, but is not limited to, ginseng (Panax ginseng), red ginseng, or wild ginseng.

[0049] The above ginseng seed oil (GSO) has been attracting attention in recent years as a source of new bioactive ingredients with various potential applications in the health and wellness fields, and may be used interchangeably with another term, ginseng seed oil.

[0050] The above ginseng seeds can be used without limitation, whether cultivated or commercially available, and can be used as is after washing or after drying. All drying methods, including sun drying, shade drying, hot air drying, and natural drying, can be used.

[0051] In one specific example of the present invention, ginseng seed oil or ginseng seed oil may refer to oil extracted by extracting ginseng seeds or seeds using a specific extraction method.

[0052] In another specific embodiment of the present invention, ginseng seed oil or ginseng seed oil may refer to oil extracted by separating ginseng seeds or seeds into endosperm and husk and then extracting them using a known extraction method.

[0053] In the present invention, the extraction means a preparation obtained by squeezing an extract from a raw material and evaporating the extract to concentrate it, and may be an extract obtained by extraction treatment, a diluted or concentrated extract, a dried product obtained by drying the extract, a controlled substance or purified product thereof.

[0054] In addition, various methods such as hot water extraction, immersion extraction, reflux extraction, reflux cooling extraction, solvent extraction, steam distillation, supercritical extraction, ultrasonic extraction, dissolution, and pressing can be used as extraction methods, but are not limited thereto, and preferably, the extraction can be performed using supercritical extraction.

[0055] The above ginseng seed oil or wild ginseng seed oil can be obtained in a liquid form by extraction, heating, and / or filtering at room temperature using conventional methods known in the art, or can be further subjected to solvent evaporation, spray drying, or freeze drying. In addition, the above ginseng seed oil can be manufactured into a powder form through additional processes such as reduced pressure distillation and freeze drying or spray drying. In addition, the oil can also be obtained as a fraction further purified using various chromatography methods such as silica gel column chromatography, thin layer chromatography, and high performance liquid chromatography.

[0056] The extraction temperature of the above ginseng seeds or ginseng seed oil is preferably 10 to 100°C, specifically 30 to 60°C, and the extraction time is not particularly limited, but may be 1 to 20 hours, specifically 5 to 12 hours, and more specifically 7 to 10 hours.

[0057] In a specific embodiment of the present invention, the ginseng seed oil was extracted using a supercritical extraction method, and obtained by performing extraction at a pressure of 360 bar and a temperature of 50°C for 9 hours.

[0058]

[0059] The composition containing the above ginseng seed oil can be used in the form of a nanoemulsion (GSO-nanoemulsion, GSO-NE) of ginseng seed oil, and at this time, the nanoemulsion of ginseng seed oil can be formed or formulated as an oil-in-water (o / w) nanoemulsion.

[0060] In the present invention, an emulsion refers to a mixed phase in which at least one immiscible liquid, either an oil phase or an aqueous phase, is dispersed in the form of fine particles (dispersion medium) in another liquid (dispersion medium). Emulsions are typically classified into macroemulsions, microemulsions, and nanoemulsions depending on the particle size of the dispersed phase. At this time, the stability, rheological properties, texture, etc. of the emulsion are determined by the physical properties of each component of the oil / water / surfactant system constituting the emulsion, the miscibility between each component, and the tension of the water / oil interface.

[0061] The above ginseng seed oil water-in-oil nanoemulsion may further contain a surfactant.

[0062] In the present invention, the term "surfactant" refers to a substance that has both a polar (hydrophilic) portion and a non-polar (lipophilic / hydrophobic) portion. Water and oil do not originally mix well and form a boundary, but when a surfactant is added, the boundary is activated and the substance is mixed.

[0063] The above surfactant may be an amphiphilic surfactant, a hydrophobic surfactant, or a combination thereof.

[0064] In the present invention, the term "amphiphilic surfactant" refers to a surfactant in which one molecule simultaneously has a hydrophilic (polar) portion and a hydrophobic (non-polar) portion, and although not particularly limited, the amphiphilic surfactant may include Tween 80.

[0065] In the present invention, the term "hydrophobic surfactant" means a surfactant having hydrophobic, i.e. nonpolar, properties, and the hydrophobic surfactant is not particularly limited, but may preferably include Span 80.

[0066] In one specific embodiment of the present invention, the surfactant may include a mixture of Tween 80 and Span 80, and more specifically, may be mixed in an amount of 2 to 3 wt% of Tween 80 and 2 to 3 wt% of Span 80 relative to the total composition, but is not limited thereto.

[0067] In addition, the amphiphilic surfactant and hydrophobic surfactant can be mixed with ginseng seed oil in a weight ratio of 1:1 to 1:4, and specifically, can be mixed in a weight ratio of 1:2.

[0068] The above ginseng seed oil-in-water nanoemulsion may be characterized in that the nanoemulsion is manufactured using an ultrasonic treatment time of 15 to 25 minutes and at an ultrasonic treatment amplitude of 50 to 70%.

[0069] In a specific example of the present invention, the ginseng seed oil-in-water ginseng seed oil and surfactant can be prepared by mixing them and then ultrasonicating them, and more specifically, it can be prepared by mixing 5 to 20 wt% of ginseng seed oil, 1 to 5 wt% of a surfactant mixture, and 75 to 90 wt% of water and then ultrasonicating them.

[0070] In a specific embodiment of the present invention, the water-in-oil nanoemulsion may have a water particle size of 1 to 600 nm, more specifically 100 to 500 nm, and even more specifically 200 to 400 nm at room temperature conditions.

[0071] In a specific embodiment of the present invention, the oil-in-water nanoemulsion may have a zeta potential of -10 to -50 mV, more specifically -20 to -40 mV, and most specifically -35 to -37 mV.

[0072] In addition, the above ginseng seed oil oil-in-water nanoemulsion may be characterized by a reduced droplet size and an improved zeta potential compared to nanoemulsions formulated under other conditions, thereby significantly improving colloidal stability.

[0073] In a specific embodiment of the present invention, when the amphiphilic surfactant and the hydrophobic surfactant were mixed with ginseng seed oil in a weight ratio of 1:1 to 1:4, it was confirmed that the most optimized formulation was obtained when the mixture was mixed in a weight ratio of 1:2, and further, when the ultrasonic treatment time was 15 to 25 minutes and the ultrasonic treatment amplitude was 50 to 70%, it was confirmed that the droplet size was reduced and the zeta potential was improved compared to nanoemulsions formulated under other conditions, thereby excellently improving colloidal stability.

[0074] In addition, in a specific embodiment of the present invention, it was confirmed that the GSO-NE of the present invention was the most stable, showing the lowest zeta potential (-36.68 mV) at a mixing ratio of 2:1, which is 10 wt% of GSO and 5 wt% of surfactant among various GSO concentrations, and that the GSO-nanoemulsion of the present invention had optimal characteristics at 20 minutes of ultrasonic treatment and 60% ultrasonic amplitude, with the droplet size reduced to 359.6 nm and the zeta potential improved to -37 mV, thereby confirming that the colloidal stability was excellently improved compared to the initial or other conditions.

[0075] In particular, in a specific embodiment of the present invention, it was confirmed that the GSO-NE of the present invention effectively inhibits coronavirus by reversing the expression of the ABCG transporter suppressed by coronavirus and increasing it again, i.e., reactivating it.

[0076]

[0077] Another aspect of the present invention provides an antiviral composition against coronavirus comprising an ABCG transporter activator comprising ginseng seed oil.

[0078] The above terms, “ginseng seed oil”, “ABCG transporter activator”, etc., are as described above.

[0079] In the present invention, "antiviral" refers to, but is not limited to, the effect of inhibiting the proliferation or replication of pathogenic viruses, thereby reducing, suppressing, or preventing viral infection. "Pathogenic viruses" whose proliferation or replication is inhibited by the antiviral activity are not limited thereto.

[0080] An example of this is a virus that causes disease in animals or humans, specifically the virus may be a coronavirus, and more specifically one or more selected from the group consisting of SARS-CoV-2 (Covid 19), SARS-CoV, MERS-CoV and human coronavirus OC43 (HCoV-OC43), and even more specifically human coronavirus OC43 (HCoV-OC43).

[0081] The term "SARS-CoV-2" in the present invention refers to a viral strain that causes coronavirus disease 2019 (COVID-19), a respiratory illness. As announced by the National Institutes of Health (NIH), it is the successor of SARS-CoV. SARS-CoV-2 is a positive-sense single-stranded RNA virus. It is contagious in humans, and the World Health Organization (WHO) has declared the ongoing pandemic of COVID-19 a Public Health Emergency of International Concern (PHEIC).

[0082] Taxonomically, SARS-CoV-2 is a strain of SARS-CoV, is thought to have zoonotic origins, and shares close genetic similarities with bat coronaviruses. SARS-CoV-2 is primarily transmitted through close person-to-person contact and / or through droplets produced when coughing or sneezing, and enters human cells primarily by binding to the angiotensin-converting enzyme 2 (ACE2) receptor.

[0083] The term "SARS-CoV" in the present invention is an ssRNA virus belonging to the genus Betacoronavirus with an envelope. Its entire genome consists of 29,727 nucleotides, making it the virus with the largest genome among RNA viruses known to date. The genome of SARS-CoV is known to have 11 open reading frames (ORFs) and encode 23 types of proteins. The main structural proteins of SARS-CoV have been revealed to be nucleocapsid (N), spike (S), membrane (M), and small envelope (E) proteins.

[0084] Sequence analysis of these proteins revealed that SARS-CoV has a very low sequence homology of approximately 40-50% with other coronaviruses. According to phylogenetic classification based on antigenicity of coronaviruses, SARS-CoV belongs to lineage B, which includes bat SARS-like coronaviruses and other bat-derived coronaviruses, while MERS-CoV belongs to lineage C, which includes bat-derived coronaviruses (Trends Microbiol 24:490-502). Furthermore, human coronavirus-OC43 belongs to lineage D, which is formed by recombination between lineage B and lineage C.

[0085] The term "MERS-CoV" in the present invention is a ssRNA virus belonging to the genus Betacoronavirus newly discovered on September 24, 2012, having a structure of 5'-replicase-spikeenvelope-membrane-nucleocapsid-poly(A)-3'[5''-ORF1a / bSEMN-poly(A)]. The MERS-CoV genome is phylogenetically classified into clade A and clade B, with the initial MERS cases being clade A (EMC / 2012 and Jordan-N3 / 2012), and the newly reported cases being genetically distinct clade B (Emerging Infectious Diseases, Vol. 20, No. 6, June 2014).

[0086] The term "human coronavirus OC43 (HCoV-OC43)" used in the present invention is known to cause the common cold. Among them, HCoV-OC43 is the most common human coronavirus and is known to cause upper respiratory tract infection. Recent studies have shown that many children seroconvert to HCoV early before the age of 2, with HCoV-OC43 being the most common. HCoV-OC43 infection has been shown to be involved in the development of severe respiratory disease in children or the elderly, and has been confirmed to cause severe lower respiratory disease in children.

[0087] The composition may be characterized by having antiviral activity against coronaviruses by increasing or reactivating the expression of ABCG transporters suppressed by coronavirus infection, thereby reactivating the inactivated pathway.

[0088] Additionally, the composition may be characterized by having anti-inflammatory activity together with or simultaneously with antiviral activity.

[0089]

[0090] The above composition may additionally contain a substance having antiviral activity, and specifically, may additionally contain silydianin.

[0091] In the present invention, silydianin is a type of flavolignan that forms a silymarin complex together with silybin, isosilybin, and silychristin.

[0092] For example, the above oil-in-water nanoemulsion can be usefully utilized because it has antiviral properties when combined with silydianin.

[0093] In one specific example of the present invention, it was confirmed that the GSO-NE of the present invention effectively inhibits coronavirus by reversing and reactivating the expression of the ABCG transporter, which has been suppressed by coronavirus. Accordingly, the antiviral activity and the therapeutic, preventive, and ameliorating effects of viral infectious diseases were confirmed by administering the composition of the present invention.

[0094]

[0095] Another aspect of the present invention provides a pharmaceutical composition for treating or preventing a coronavirus infectious disease, comprising an ABCG transporter activator including the ginseng seed oil.

[0096] The above terms, “ginseng seed oil,” “ABCG transporter activator,” “coronavirus,” etc., are as described above.

[0097] The term "viral infectious disease" of the present invention refers to a disease caused by infection with a virus. The term "infection" refers to a state in which a pathogenic microorganism invades the body of a host organism and grows and proliferates. The viral infectious disease of the present invention may specifically be an infectious disease caused by infection with a coronavirus, and more specifically, may be one or more infectious diseases selected from the group consisting of SARS-CoV-2 (Covid 19), SARS-CoV-2, MERS-CoV-2, and human coronavirus OC43, and more specifically, may be an infectious disease caused by human coronavirus OC43.

[0098] The pharmaceutical composition of the present invention has been confirmed to effectively inhibit coronavirus by reversing the expression of the ABCG transporter suppressed by coronavirus and increasing it again, i.e., reactivating it. Furthermore, it has been confirmed to have an excellent effect in suppressing the inflammatory response caused by coronavirus infection. Therefore, it can be usefully used for the treatment, prevention, and improvement of viral infection diseases.

[0099] The term "prevention" in the present invention means any act of inhibiting or delaying a coronavirus infectious disease by administering the pharmaceutical composition.

[0100] In addition, the term "treatment" of the present invention means any act of improving or completely curing the symptoms of a coronavirus infection disease by administering the pharmaceutical composition.

[0101] The pharmaceutical composition may comprise a pharmaceutically acceptable carrier.

[0102] The term "pharmaceutically acceptable carrier" of the present invention may refer to a carrier, excipient, or diluent that does not stimulate a living organism and does not inhibit the biological activity and properties of the compound to be injected, and specifically, may be a non-naturally occurring carrier. The type of the carrier usable in the present invention is not particularly limited, and any pharmaceutically acceptable carrier commonly used in the art may be used. Non-limiting examples of the carrier include saline solution, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and the like. These may be used alone or in combination of two or more.

[0103] The pharmaceutical composition comprising a pharmaceutically acceptable carrier may be administered orally or parenterally in various dosage forms. When formulated, it is prepared using commonly used diluents or excipients, such as fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants.

[0104] Specifically, solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations can be prepared by mixing the compound with at least one excipient, such as starch, calcium carbonate, sucrose, lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Liquid preparations for oral administration include suspensions, oral solutions, emulsions, syrups, etc., and in addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc. can be included. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspending agents can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases can include withepsol, macrogol, Tween 61, cocoa butter, laurin butter, and glycerogelatin.

[0105] The above pharmaceutical composition can be administered in a pharmaceutically effective amount.

[0106] The above "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the effective dosage level can be determined according to the type and severity of the individual, age, sex, type of infected virus, activity of the drug, sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, concurrently used drugs, and other factors well known in the medical field. For example, the pharmaceutical composition may be administered such that the ginseng seed oil oil-in-water nanoemulsion of the present invention or the composition comprising silydianin therewith is administered at 0.0001 to 1,000 mg / kg per day, specifically 0.001 to 100 mg / kg.

[0107] The above administration means introducing the composition of the present invention into a patient by any suitable method, and the route of administration of the composition may be any common route as long as it can reach the target tissue. It may be intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration, or intranasal administration, but is not limited thereto.

[0108] The pharmaceutical composition of the present invention may be administered daily or intermittently, and may be administered once a day or divided into 2-3 doses. When the two active ingredients are each administered as single agents, the doses may be the same or different. Furthermore, the composition of the present invention may be used alone or in combination with other drug treatments for the prevention or treatment of cancer. Taking all of the above factors into consideration, it is important to administer an amount that achieves maximum effect with the minimum amount without side effects, and this can be readily determined by those skilled in the art.

[0109] The above-mentioned entity refers to any animal, including rats, mice, and livestock, including humans, that has developed or may develop a coronavirus infectious disease. Specifically, it may be a mammal, including humans.

[0110] In a specific embodiment of the present invention, it was confirmed that the GSO-NE of the present invention effectively inhibits coronavirus by reversing and reactivating the expression of the ABCG transporter, which has been suppressed by coronavirus. Accordingly, the antiviral activity and the therapeutic, preventive, and ameliorating effects of viral infectious diseases were confirmed by administering the composition of the present invention.

[0111] Through this, it can be seen that a composition including an ABCG transporter activator including a ginseng seed oil-in-water nanoemulsion has an antiviral effect against coronavirus and an effect of preventing, improving, and treating diseases caused by coronavirus infection.

[0112]

[0113] Another aspect of the present invention provides a pharmaceutical composition for preventing or improving coronavirus infectious disease, comprising an ABCG transporter activator comprising a ginseng seed oil-in-water nanoemulsion of the present invention.

[0114] The above terms, “ginseng seed oil,” “ABCG transporter activator,” “coronavirus infectious disease,” “prevention,” etc., are as described above.

[0115] The term “improvement” as used above means any act of improving or curing the symptoms of a coronavirus infection disease by administering the composition.

[0116]

[0117] The term "quasi-drug" used in the present invention means an article other than a device, machine or apparatus used for the purpose of diagnosing, treating, alleviating, managing or preventing a disease in humans or animals, and an article other than a device, machine or apparatus used for the purpose of exerting a pharmacological effect on the structure and function of humans or animals.

[0118] In the present invention, the above-mentioned pharmaceutical composition may have an effect of preventing or improving a viral infectious disease, but is not limited thereto.

[0119] In addition to the above-mentioned ingredients, the quasi-drug composition of the present invention may further include a pharmaceutically acceptable carrier, excipient, or diluent, as needed. The pharmaceutically acceptable carrier, excipient, or diluent is not limited as long as it does not impair the effects of the present invention, and may include, for example, fillers, bulking agents, binders, wetting agents, disintegrants, surfactants, lubricants, sweeteners, fragrances, preservatives, etc.

[0120]

[0121] Another aspect of the present invention provides a food composition for preventing or improving coronavirus infectious disease, comprising an ABCG transporter activator comprising a ginseng seed oil-in-water nanoemulsion of the present invention.

[0122] The above terms, “ginseng seed oil,” “ABCG transporter activator,” “coronavirus infectious disease,” “prevention,” “improvement,” etc., are as described above.

[0123] The term "food" of the present invention includes dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, beverages, tea, drinks, alcoholic beverages, vitamin complexes, health functional foods, and health foods, and includes all foods in the conventional sense.

[0124] The above functional health food is the same term as food for special health use (FoSHU), and refers to a food with high medical and healthcare effects that is processed to efficiently exhibit a bioregulatory function in addition to providing nutrition. Here, "functionality" means regulating nutrients for the structure and function of the human body or obtaining a useful effect for health purposes such as physiological action. The food of the present invention can be manufactured by a method commonly used in the art, and during the manufacturing process, raw materials and ingredients commonly added in the art can be added. In addition, the formulation of the food can be manufactured without limitation as long as it is a formulation recognized as a food. The food composition of the present invention can be manufactured in various forms of formulations, and unlike general drugs, it has the advantage of not having side effects that may occur when taking drugs for a long time by using food as a raw material, and is highly portable, so the food of the present invention can be consumed as a supplement to enhance the effect of preventing or improving inflammatory diseases.

[0125] The above-mentioned "health food" refers to foods that have a more active health maintenance or promotion effect than regular foods, while "health supplement food" refers to foods intended for health supplementation. In some cases, the terms "health functional food," "health food," and "health supplement food" are used interchangeably.

[0126] Specifically, the health functional food means a food product manufactured by adding the compound of the present invention to food materials such as beverages, teas, spices, gums, and confectionery, or by manufacturing it in the form of encapsulation, powder, suspension, etc., and which has a specific health effect when consumed, but unlike general drugs, it has the advantage of not having side effects that may occur with long-term use of drugs made from food as a raw material.

[0127] The food composition of the present invention can be used very usefully because it can be consumed on a daily basis and is expected to be highly effective in preventing or improving cancer.

[0128] The above food composition may additionally include a physiologically acceptable carrier. The type of the carrier is not particularly limited, and any carrier commonly used in the art may be used.

[0129] In addition, the food composition may include additional ingredients commonly used in food compositions to improve odor, taste, sight, etc. For example, it may include vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantothenic acid, etc. In addition, it may include minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), chromium (Cr), etc. In addition, it may include amino acids such as lysine, tryptophan, cysteine, and valine.

[0130] In addition, the food composition may include food additives such as preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), sterilizers (bleaching powder and high-purity bleaching powder, sodium hypochlorite, etc.), antioxidants (butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), etc.), colorants (tar colorants, etc.), color developers (sodium nitrite, sodium nitrite, etc.), bleaching agents (sodium sulfite), seasonings (MSG, monosodium glutamate, etc.), sweeteners (dulcin, cyclamate, saccharin, sodium, etc.), flavorings (vanillin, lactones, etc.), leavening agents (alum, D-potassium hydrogen tartrate, etc.), reinforcing agents, emulsifiers, thickeners (pastes), film agents, gum bases, foaming agents, solvents, and improvers. The above additives can be selected according to the type of food and used in an appropriate amount.

[0131] The oil of the present invention can be added as is or used together with other foods or food ingredients, and can be used appropriately according to a conventional method. The amount of the active ingredient mixed can be appropriately determined depending on its purpose of use (prevention, health, or therapeutic treatment). Generally, when manufacturing a food or beverage, the food composition of the present invention can be added to the food or beverage in an amount of 50 parts by weight or less, specifically 20 parts by weight or less. However, when consumed for a long period of time for health and hygiene purposes, the content below the above range can be included, and since there is no problem in terms of safety, the active ingredient can also be used in an amount above the above range.

[0132] As an example, the food composition of the present invention can be used as a health beverage composition, and in this case, it can contain various flavoring agents or natural carbohydrates as additional ingredients like a regular beverage. The above-mentioned natural carbohydrates can be monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; sugar alcohols such as xylitol, sorbitol, and erythritol. The sweetener can be a natural sweetener such as thaumatin and stevia extract; a synthetic sweetener such as saccharin and aspartame, etc. The proportion of the natural carbohydrate can be generally about 0.01 to 0.04 g, specifically about 0.02 to 0.03 g per 100 mL of the health beverage composition of the present invention.

[0133] In addition to the above, the health beverage composition may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, or carbonating agents. In addition, it may contain fruit pulp for the production of natural fruit juice, fruit juice drinks, or vegetable drinks. These ingredients may be used independently or in combination. The proportion of these additives is not particularly important, but is typically selected within the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the health beverage composition of the present invention.

[0134] The food composition of the present invention may contain various weight % if it can exhibit the effect of preventing or improving a viral infection disease, but specifically, the ginseng gold nanoparticles and silidianin of the present invention may contain 0.00001 to 100 wt % or 0.01 to 80 wt % relative to the total weight of the food composition, but is not limited thereto.

[0135]

[0136] Another aspect of the present invention provides a feed composition comprising an ABCG transporter activator including ginseng seed oil of the present invention.

[0137] The above terms, “ginseng seed oil,” “ABCG transporter activator,” “coronavirus infectious disease,” “prevention,” “improvement,” etc., are as described above.

[0138] The term "feed" in the present invention means any natural or artificial diet, meal, etc., or ingredients of said meal, intended for or suitable for an animal to eat, ingest, or digest. The type of said feed is not particularly limited, and feed commonly used in the relevant technical field can be used. Non-limiting examples of said feed include plant feeds such as grains, roots, fruits, food processing by-products, algae, fibers, pharmaceutical by-products, oils, starches, meal, or grain by-products; and animal feeds such as proteins, inorganic substances, oils, minerals, oils, single-cell proteins, zooplankton, or food. These may be used alone or in combination of two or more.

[0139]

[0140] Another aspect of the present invention provides a pharmaceutical composition for preventing or treating an inflammatory disease caused by a coronavirus, comprising an ABCG transporter activator including ginseng seed oil of the present invention.

[0141] The above terms, “ginseng seed oil”, “ABCG transporter activator”, “coronavirus”, “prevention”, “improvement”, “pharmaceutical composition”, etc., are as described above.

[0142] The above inflammatory disease is a disease caused by inflammation that appears as an internal response to damage or infection of a specific tissue, and specifically refers to a disease caused by inflammation induced by a coronavirus, and more specifically, may be, but is not limited to, pneumonia induced by a coronavirus, systemic inflammation induced by LPS, or multisystem inflammatory syndrome.

[0143] The above pneumonia is an inflammation of the lungs caused by a coronavirus infection, and may include pulmonary symptoms caused by disruption of the normal function of the lungs, as well as systemic symptoms throughout the body, such as coughing due to respiratory irritation, phlegm due to discharge of inflammatory substances, and difficulty breathing due to disruption of breathing function. In addition, it may be a disease that includes digestive symptoms such as nausea, vomiting, and diarrhea, systemic symptoms such as headache, fatigue, muscle pain, and joint pain, fever, chills, etc.

[0144] The above multisystem inflammatory syndrome is a disease in which an inflammatory response occurs throughout the body, and is also called multisystem inflammatory syndrome in children (MIS-C). It is a disease presumed to be related to COVID-19, and is reported to present with symptoms such as high fever, skin rash, and in severe cases, toxic shock accompanied by inflammation of the coronary artery.

[0145]

[0146] Another aspect of the present invention provides an anti-inflammatory composition comprising an ABCG transporter activator including the ginseng seed oil of the present invention.

[0147] The above terms, “ginseng seed oil,” “ABCG transporter activator,” “anti-inflammation,” etc., are as described above.

[0148]

[0149] Another aspect of the present invention provides a food composition for preventing or improving an inflammatory disease caused by a coronavirus, comprising an ABCG transporter activator including ginseng seed oil of the present invention.

[0150] The above terms, “ginseng seed oil,” “ABCG transporter activator,” “inflammatory disease caused by coronavirus,” “prevention,” “improvement,” “food composition,” etc., are as described above.

[0151]

[0152] Another aspect of the present invention provides a pharmaceutical composition for preventing or improving an inflammatory disease caused by a coronavirus, comprising an ABCG transporter activator including ginseng seed oil of the present invention.

[0153] Another aspect of the present invention provides a pharmaceutical feed composition for preventing or improving inflammatory diseases caused by coronavirus, comprising an ABCG transporter activator including ginseng seed oil of the present invention.

[0154] The above terms, “ginseng seed oil,” “ABCG transporter activator,” “inflammatory disease caused by coronavirus,” “prevention,” “improvement,” “quasi-drug composition,” “feed composition,” etc., are as described above.

[0155]

[0156] Another aspect of the present invention provides a method for preparing an antiviral composition for coronavirus comprising an ABCG transporter activator comprising a ginseng seed oil oil-in-water nanoemulsion, the method comprising the step of formulating the ginseng seed oil of the present invention into an oil-in-water nanoemulsion.

[0157] The above terms, “ginseng seed oil”, “oil-in-water nanoemulsion”, “formulation”, “ABCG transporter activator”, “coronavirus”, “antiviral composition”, etc., are as described above.

[0158] The above-mentioned formulating step is a step of preparing the prepared ginseng seed oil into an oil-in-water nanoemulsion formulation, wherein the ginseng seed oil can be prepared by drying, crushing, and extracting ginseng seeds. The step of preparing the ginseng seed oil and then formulating it into an oil-in-water nanoemulsion of the ginseng seed oil can mean formulating the ginseng seed oil by adding a mixture of an amphiphilic surfactant and a hydrophobic surfactant to the ginseng seed oil, and the mixture of the amphiphilic surfactant and the hydrophobic surfactant can be formulated by mixing it with the ginseng seed oil in a ratio of 1:1 to 1:4, and specifically, it is preferably mixed in a ratio of 1:2.

[0159] Hereinafter, embodiments of the present invention will be described in detail with reference to the drawings so that those skilled in the art can easily implement the present invention. Unless otherwise defined, terms used herein should be interpreted as generally understood by those skilled in the art.

[0160] The drawings and examples of this specification are provided to enable those skilled in the art to easily understand and practice the present invention. Contents that may obscure the gist of the invention may be omitted from the drawings and examples, and the present invention is not limited to the drawings and examples.

[0161] Throughout this specification, "%" used to indicate the concentration of a particular substance is (wt / wt)% for solid / solid, (wt / vol)% for solid / liquid, and (vol / vol)% for liquid / liquid, unless otherwise stated.

[0162] Unless otherwise specified, all numbers, values ​​and / or expressions expressing ingredients, reaction conditions and quantities of ingredients used herein are to be understood as being modified in all instances by the term "about" because these numbers are approximations that inherently reflect, among other things, the various uncertainties of measurement encountered in obtaining such values.

[0163] And, when a numerical range is disclosed in this specification, such range is continuous and includes all values ​​from the minimum value to the maximum value inclusive, unless otherwise specified.

[0164] Additionally, the term "or" in this specification is intended to mean an inclusive "or" rather than an exclusive "or." That is, where a connection or use between components is not otherwise specified or clear from context, i.e., if X includes A; X includes B; or X includes both A and B, "X includes A or B" can apply to any of these cases.

[0165] Throughout this specification, whenever a part is said to "include" a component, this means that it may include other components, but not to the exclusion of other components, unless specifically stated otherwise.

[0166]

[0167] Hereinafter, the present invention will be described in more detail through examples. These examples are intended to more specifically illustrate the present invention, and the scope of the present invention is not limited to these examples.

[0168]

[0169] Experimental Example 1. Extraction and Physicochemical Analysis of Ginseng Seed Oil (GSO)

[0170] Ginseng (Panax ginseng CAMeyer) seeds aged 4-5 years were dried and ground. Subsequently, ginseng seed oil (GSO) was produced by supercritical fluid extraction (SFE) of the ginseng seeds using a PBKSFE100LL_SEMI device. The SFE process was performed at 360 bar, 50°C, and for 9 hours.

[0171] The above-mentioned ginseng seed oil (GSO) was analyzed for various physicochemical parameters according to the standard methods of the Association of Official Analytical Chemists (AOAC-1990). The physicochemical parameters are summarized as follows.

[0172]

[0173] 1-1. Acid value

[0174] 2.5 g of the GSO prepared in Example 1 was weighed and placed in a conical flask, and 5 cm3 of chloroform and 25 cm3 of diethyl ether and ethanol were mixed in a 1:1 (v / v) ratio and added. A small amount of phenolphthalein indicator was added to the mixture, and titration was performed with a standardized 0.1 M potassium hydroxide (KOH) solution. The titration endpoint was set when a pink color appeared and persisted for at least 30 seconds.

[0175]

[0176] 1-2. Saponification value

[0177] 2.5 g of the GSO prepared in Example 1 was placed in a conical flask, and the oil sample was dissolved in 5 cm3 of chloroform. 25 cm3 of a 0.5 M KOH solution was then added to the flask, the inlet of the flask was corked, and the mixture was refluxed for 30 minutes to promote the saponification process. After reflux, the mixture was transferred to another conical flask, and a few drops of phenolphthalein indicator were added. The resulting mixture was titrated with a 0.5 M hydrochloric acid (HCl) solution until the pink color disappeared.

[0178]

[0179] 1-3. Iodine value

[0180] Using a 100 cm3 glass stoppered flask, 0.5 g of GSO in oil form was dissolved in 10 cm3 of chloroform. Next, 25 cm3 of Wij's solution was added to the flask, left in a dark place for 30 minutes, and 20 cm3 of a 10% potassium iodide (KI) solution was added. To confirm the end point of the titration in the resulting solution, a standardized 0.1 M sodium sulfate (Na2S2O3) solution was added.

[0181]

[0182] 1-4. Specific gravity

[0183] The initial weight (W0) was obtained by measuring the weight of a clean and dry density bottle with a capacity of 25 ㎤, GSO was put into the density bottle, the bottle was corked, and the weight (W1) was measured again. Then, the density bottle was filled with distilled water, the weight (W2) was measured, and the specific gravity of the oil was calculated by comparing the weights of the density bottle filled with oil and water.

[0184]

[0185] 1-5. Refractive index

[0186] An Abbe's refractometer was used to measure the refractive index. Specifically, a small amount of GSO was first transferred to the glass slide of the refractometer, and water at 30°C was circulated around the slide to maintain a constant temperature. The dark area confirmed by the refractometer was adjusted to coincide with the intersection point to avoid parallax error. The scale pointer at this time indicates the refractive index. Before sample analysis, the refractometer was calibrated using distilled water to measure the refractive index of water at a specified temperature. The measurement was repeated three times for each sample, and the average value was obtained.

[0187]

[0188] 1-6. Peroxide value

[0189] In a 250 cm3 Erlenmeyer flask, 1 g of GSO, 1 g of potassium iodide, and 20 cm3 of a mixture of glycerol acetic acid and chloroform (3:2 by volume) were mixed. The mixture was heated and boiled for 1 minute, and the boiled solution was transferred to a flask containing 20 cm3 of a 5% potassium iodide solution. Three drops of starch solution were then added to the mixture to serve as an indicator. In addition, a titration with standardized 0.025 N sodium sulfate (Na2S2O3) was performed to determine the end point of the titration in the solution.

[0190]

[0191] Experimental Example 2. GSO Lipid Grade Analysis

[0192] 2-1. Free fatty acids

[0193] First, GSO (3.5 g) prepared in Example 1 was transferred to a clean, dry flask containing 225 mL of neutralized alcohol. The resulting mixture was titrated with a 0.25 N sodium hydroxide (NaOH) solution until a light pink color appeared. In addition, the volume of 0.25 N NaOH required for titration with a blank solution was measured.

[0194]

[0195] 2-2. Phytosterols

[0196] The identification and quantification of phytosterols contained in the GSO manufactured in Example 1 were confirmed through HPLC equipped with a UV detector. The analysis was performed using an Agilent 1260 infinity Ⅱ Quat Pump (California, USA) equipped with a variable wavelength (VW) detector. Specifically, an INNO C18 column with a 6X150 mm diameter and a 3 μm particle size was used, and a constant temperature of 28 °C was maintained. Methanol (MeOH) configured as an isoctahedral system was used as the mobile phase, and 10 μL was used. The flow rate was set to 0.95 mL / min, and detection was performed at a wavelength of UV 205 nm. This HPLC analysis method accurately identified and quantified phytosterols present in GSO.

[0197]

[0198] 2-3. Oxidation stability

[0199] 2-3-1. Analysis of conjugated dienes (CD) and conjugated trienes (CT)

[0200] GSO (0.05 g) prepared in Example 1 was dissolved in 10 mL of isooctane, and absorbance measurements were performed using a Shimadzu UV-1800 spectrophotometer (Shimadzu Europe GmbH, Germany). The absorbance values ​​were recorded at wavelengths of 232 nm and 270 nm when the spectral scan was completed. These absorbance values ​​were also used to calculate the concentrations of primary and secondary oxidation products.

[0201]

[0202] 2-3-2. Oxidation stability

[0203] To determine oxidation stability, a Metrohm Rancimat instrument (model 743, Herisau, Switzerland) was used. Specifically, approximately 3 g of GSO was weighed into the reaction vessel, and 70 mL of deionized water was added to the conductivity cell. Clean, filtered, dry air was then bubbled through the hot oil at a temperature of 110°C at a flow rate of 20 L / h.

[0204]

[0205] 2-4. GSO fatty acid analysis

[0206] For the analysis of fatty acids contained in GSO, gas chromatography (GC) was performed using a capillary GC column (SP-2560; 100 mX0.25 mm inner diameter, 0.20 μm film). The analysis was performed using an Agilent 7890A GC system (Agilent Technologies, Santa Clara, CA, USA) with a flame ionization detector, using helium as a carrier gas at a velocity of 20 cm / s.

[0207]

[0208] Experimental Example 3: Synthesis and Characterization of GSO-NE

[0209] In GSO, the oil phase was composed of various proportions, ranging from 5 to 20%. Tween80 (HLB15) and Span80 (HLB4.3), as shown in Table 4 below, were also used in the nanoemulsion synthesis process. Specifically, after Tween80 / Span80 were thoroughly mixed with the oil phase, the aqueous phase was stirred constantly at 500 rpm / min for 15 min. The resulting mixture exhibited a consistent milky white color. Subsequently, ultrasonication was performed using a probe sonicator (Hielscher UP 200st with tip size S26d2, Germany). Ultrasonication was performed within defined parameters, with the amplitude adjusted from 20 to 80% for 5 to 40 min. To evaluate the stability at various temperatures, the nanoemulsions prepared above were analyzed at temperatures ranging from 4 to 37°C. The cold conditions were set using ice, and the temperature control, oxidation prevention, minimization of heat influence, consistency and reproducibility were checked. The average particle size and homogeneity were checked using a Malvern Zeta Nano S 90 device (Malvern Panalytical Ltd, UK), and the analysis was performed using intensity-based calculations in terms of both the number and mass of particles to calculate the average particle size. In addition, the polydispersity index (PDI) was calculated to check the uniformity of the manufactured formulation, and Fourier-transform infrared (FTIR, PerkinElmer, Waltham, MA, USA) spectroscopy in the range of 4,000-500 cm -1 4 cm within range -1 It was operated at a resolution of . This method was used to identify and classify peak shifts in the functional groups of GSO and GSO-NE.

[0210]

[0211] Experimental Example 4. Cell Viability Analysis in Vero E6 Cells

[0212] Vero E6 cells derived from African green monkey kidney (ATCC ® CRL-1586) was cultured in Dulbecco's modified Eagle's medium (Walkersville, MD, USA). The medium was supplemented with 10% FBS (Gibco, Life Technologies, Grand Island, NY, USA), 1% 10 mM HEPES / 0.85 NaCl (Lonza, BioWhittaker ® , MD, Walkersville), 100 U / mL penicillin-100 μg / mL streptomycin (Gibco, Life Technologies, Grand Island, NY, USA). The cells were cultured in an environment maintained at 37°C and 5% CO2 concentration. Vero E6 cells were seeded in each well of a 96-well plate at a density of 4 × 10 4 After seeding, the cells were cultured for 24 hours, and treated with GSO-NE at various concentrations ranging from 0.625 to 40 μg / mL, and cultured for an additional 48 hours at 37°C. Then, MTT solution (10 μL) was treated to each well, and cultured for 3 hours at 37°C, and then 100 μL of DMSO was additionally added to each well, and then the absorbance was measured at a wavelength of 595 nm to confirm the cell viability.

[0213]

[0214] Experimental Example 5. Antiviral Activity Analysis against HCoV-OC43

[0215] Human coronavirus OC43 (HCoV-OC43, KBPV-VR-8) was purchased from the Korean Virus Bank (Seoul, Korea). Vero E6 cells were then cultured in 6-well plates (Corning ® ) in 1×10 6After seeding in an amount of 10 μg / mL, the cells were cultured overnight at 37°C and 5% CO2. The cells were then washed twice with PBS and infected with HCoV-OC43 at a multiplicities of infectivity (MOI) of 0.02 for 2 hours. The plates were then manually shaken every 15-20 minutes to ensure even distribution of HCoV-OC43, and after virus absorption, the cells were treated with various concentrations of the GSO-NE of the present invention. After 48 hours post-infection, the cells and supernatant were harvested, and the degree of infection was confirmed by observing cell morphology using a microscope.

[0216]

[0217] Experimental Example 6. Transcriptome analysis of GSO-NE in Vero E6 cells

[0218] Vero E6 cells infected with HCoV-OC43 of the above Experimental Example 5 were treated with the GSO-NE of the present invention. RNA sequencing (RNA-Seq) was analyzed using total RNA encompassing fragmented cDNA generated using the TruSeq Stranded mRNA Library Prep Kit (Illumina, San Diego) as an RNA-Seq library.

[0219]

[0220] Experimental Example 7. Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR)

[0221] Total RNA was extracted using TRIzol (Ambion, Life Technologies, Carlsbad, CA, USA) and reverse transcribed into cDNA according to the guidelines of the High-Capacity RNA-to-cDNA kit (Thermo Fisher Scientific Baltics UAB, Vilnius, Lithuania). Specifically, cDNA was synthesized using an oligo-dT (deoxythymine) kit using 1 μg of RNA, and viral RNA was quantified using Power SYBR® Green PCR Master Mix (Applied Biosystems, Life Technologies Ltd., Woolston Warrington, UK). GAPDH (glyceraldehyde 3-phosphate dehydrogenase) was used as an endogenous control, and the details of specific primers are as follows (see Table 5).

[0222]

[0223] Experimental Example 8. Immunoblotting Analysis

[0224] Vero E6 cells were lysed with Ripa lysis buffer, and then treated with GSO-NE of the present invention at concentrations of 2.5 and 5 μg / mL to perform immunoblotting analysis. The antibodies used at this time are as shown in Table 6 (see Table 6 below).

[0225]

[0226] Experimental Example 9. Statistical Analysis

[0227] The experimental procedure was repeated three times, and the resulting data were expressed as the mean ± standard deviation. Statistical comparisons between the two groups were performed using Student's t-test, and the significance levels were set at *, p < 0.05; **, p < 0.01; and ***, p < 0.001.

[0228]

[0229] Example 1. Confirmation of physicochemical properties of GSO

[0230] The results of analyzing the physicochemical properties of the ginseng seed oil GSO of the present invention manufactured in the above experimental example 1 are as shown in Table 1 below.

[0231]

[0232]

[0233] As can be confirmed in Table 1 above, the acid value of the GSO manufactured in the present invention was confirmed to be 1.51±0.22, which is a low value between 1 and 2. The acid value reflects the quality of fatty acids, and a low value indicates long-term stability and reduces the risk of rancidity and peroxidation, which is an indicator of the purity and edibility of the oil. From this, it can be seen that the GSO of the present invention has long-term stability and edibility due to its low acidity.

[0234] Next, the saponification value refers to the average molecular weight of fatty acids present in the oil, and a high SV (saponification value) is related to a low molecular weight of fatty acids. The SV of the GSO of the present invention is 173.12±1.23 mgKOH / g, which is a high SV value, and thus, it can be seen that the GSO of the present invention is suitable for soap manufacturing, oil-based ice cream, and shampoo production.

[0235] In addition, according to the above Table 1, the iodine value (89.18±1.18) of the GSO of the present invention is less than 130, which shows once again that the GSO of the present invention is suitable for soap manufacturing or purification for food use, but on the other hand, it shows that the sensitivity to deterioration and oxidation is reduced, making it unsuitable for ink and paint production.

[0236] In addition, the specific gravity of GSO is 0.9074±0.02, which falls within the range reported for other oils such as cashew oil and sesame oil. Furthermore, the refractive index is also confirmed to be 1.4621±0.01, which falls within the range of general oils, as reported for peanut, sesame, sunflower, and palm oils. These results indicate that the GSO of the present invention possesses the characteristics of the aforementioned oil categories, and furthermore, the oil quality is guaranteed and it is edible.

[0237] Lastly, the peroxide value of the GSO of the present invention is 10.06±0.78 meq / kg, which also indicates that the GSO of the present invention is within a suitable edible range. Specifically, cottonseed oil (cotton seed oil, cooking oil, cotton seed oil) shows a peroxide value of 10.40 meq / kg, and since the commercial cooking oil standard stipulates that the peroxide value should not exceed 10 meq / kg, it can be confirmed once again that the GSO of the present invention is within the category of edible oils such as cooking oil.

[0238]

[0239] Example 2. Confirmation of lipid grade composition of GSO

[0240] The lipid grade of the GSO of the present invention was confirmed by the content of free fatty acids.

[0241] As a result, as can be confirmed in Table 1 above, the free fatty acid (FFA) content of GSO is 0.48±0.01%, which indicates that the ester portion of the oil has minimal change, indicating excellent oil quality. Generally, high-quality oil has a low FFA content, which improves the taste of refined vegetable oil, and in the food industry, frying oil with an FFA content exceeding 2% is generally discarded or supplemented with fresh oil to reduce the FFA level. The above value is much lower than 2%, and it can be seen that the GSO of the present invention is a high-quality oil with a very low FFA content.

[0242] Next, the content of plant sterols (phytosterols) in the GSO of the present invention was confirmed and shown in Table 2 below.

[0243]

[0244]

[0245] As a result, as can be confirmed in Table 2 above, the GSO of the present invention contains campesterol (8.07±0.09), stigmasterol (21.46±0.08), and beta-sitosterol (0.28±0.01), and it was confirmed that each value is within the recognized standard range of plant sterols (Table 2).

[0246]

[0247] Example 3. Confirmation of the oxidation stability of GSO

[0248] The oxidation stability of the GSO of the present invention was confirmed and is shown in Table 3 below.

[0249]

[0250]

[0251] Here, the oxidative stability was determined by the concentration of conjugated dienes (CD) and conjugated trienes (CT). Conjugated dienes are generated during the oxidation of polyunsaturated fatty acids and the thermal formation of oligomers and polymers, and this oxidation process is known to induce changes in the double bonds of unsaturated fatty acids.

[0252] As a result of checking the CD value, as can be confirmed in Table 3 above, the CD concentration confirmed in the GSO of the present invention was 4.16±0.03, and the concentration of conjugated trienes (CT) was confirmed to be 1.27±0.12 (Table 3). Considering that the formation of conjugated dienes generally proceeds faster than the formation of conjugated trienes, the GSO of the present invention was confirmed to have an induction time (h) of 22.08±0.03, which shows that it has excellent oxidation stability for evaluating the quality of oils and fats (Table 3).

[0253]

[0254] Example 4. Optimization of the formulation of a ginseng seed oil-in-water nanoemulsion (GSO-NE)

[0255] In order to confirm the antiviral effect of the GSO of the present invention, a delivery method using a nanoemulsion was applied to prepare a ginseng seed oil oil-in-water nanoemulsion (GSO-NE), and its formulation was optimized.

[0256] Specifically, nanoemulsions are formed by reducing the interfacial tension between immiscible substances using surfactants and co-surfactants, and the surfactants and co-surfactants play an important role in creating an optimal balance between GSOs.

[0257] Based on this, in order to optimize the formulation of the GSO nanoemulsion of the present invention, the mixing ratio of GSO and Tween 80 (amphiphilic surfactant) and Span 80 (hydrophobic surfactant) was varied to 1:1, 2:1, 3:1 or 4:1 as shown in Table 4 below, and the most optimal formulation for GSO-NE was confirmed.

[0258]

[0259]

[0260] Simultaneously, nanoemulsions were synthesized using various concentrations of GSO, and zeta potential was measured to screen for the most stable nanoemulsion formulation. A lower zeta potential indicates reduced electrostatic repulsion, which is important for controlling particle aggregation or charge neutralization in applications such as targeted drug delivery or specific material formulation.

[0261] As a result, as can be confirmed in Fig. 1, among various GSO concentrations, 10 wt% GSO was confirmed to be the most stable, showing the lowest zeta potential (-36.68 mV). Accordingly, it can be seen that the optimal concentration for synthesizing the GSO-NE of the present invention is 10 wt% GSO, and the optimal mixing ratio between the mixture of GSO and the surfactant (Tween 80, Span 80) is a weight ratio of 2:1.

[0262]

[0263] Example 5. Optimization of ultrasonic parameters for GSO-NE synthesis

[0264] To synthesize GSO-NE of the present invention, ultrasonic parameters were optimized, and the characteristics of GSO-NE manufactured through this were analyzed.

[0265] Specifically, the effects of various sonication times (5, 10, 20, and 40 min) and sonication amplitudes (20, 40, 60, and 80%) on the properties of GSB nanoemulsions (size distribution, zeta potential, and PDI, Fig. 3A-B) were investigated.

[0266] At the lowest settings initially checked, the nanoemulsion droplet size was 612 nm and the zeta potential was -9 mV, indicating relatively low stability and large droplet size. Thereafter, it was confirmed that the droplet size decreased significantly as both the sonication time and amplitude increased, and among them, the GSO-nanoemulsion of the present invention had the optimal characteristics at 20 minutes of sonication and 60% amplitude, with the droplet size decreasing to 359.6 nm and the zeta potential improving to -37 mV, confirming that the colloidal stability was significantly improved compared to the initial or other conditions.

[0267] In particular, the PDI value confirmed under the above optimal conditions was 0.24, thereby confirming that the GSO-NE of the present invention exhibits colloidal stability within an acceptable range.

[0268] In the case of nanoemulsions, zeta potential and Z-average are considered more important parameters than PDI, so based on this, the zeta potential and Z-average results suggest that a proper balance was achieved between obtaining a smaller droplet size and further having formulation stability by optimizing the ultrasonic parameters in the synthesis of the GSO-nanoemulsion of the present invention.

[0269]

[0270] Example 6. Long-term durability of GSO-NE

[0271] The long-term durability of the GSO-NE of the present invention was confirmed and is shown in Fig. 2 (Fig. 3C-D). Here, the long-term durability of the nanoemulsion was confirmed by stability and efficacy, and specifically, the influence of storage conditions, especially temperature changes, was confirmed. That is, the changes in the GSO-nanoemulsion of the present invention were confirmed at various storage temperatures (4°C, 25°C, and 37°C) and periods (1-6 months).

[0272] As a result, as can be confirmed in Fig. 2, the GSO nanoemulsion of the present invention showed excellent stability at 4 ℃ for the first 3 months, and the droplet size slightly increased from 340 nm to 341.52 nm, and the zeta potential also slightly decreased from 36 mV to 35 mV, confirming excellent stability. In addition, at the 6th month, the droplet size slightly increased to 345 nm and the zeta potential decreased to -33 mV, indicating a slight decrease in stability, but not a significant decrease, confirming that the stability is still maintained in an excellent state.

[0273] Next, at a storage temperature of 25 ℃, it was confirmed that the droplet size increased from 345 nm in 1 month to 385 nm in 6 months, and the zeta potential decreased from -35 mV to -31 mV. Similarly, at 37 ℃, it was confirmed that the droplet size of the nanoemulsion increased from 349 nm in 1 month to 392 nm in 6 months, and the zeta potential decreased from -32 mV to -29 mV.

[0274] Since the PDI value is also an indicator of the stability of GSO-NE, this was also confirmed. As a result, at 4℃, the PDI was 0.23 after 1 month and slightly decreased to 0.20 after 6 months, and the initial PDI value of the nanoemulsion stored at 25℃ was 0.21 after 1 month and slightly decreased to 0.19 after 6 months. At 37℃, it was 0.20 after 1 month and decreased to 0.16 after 6 months. That is, although the PDI value of the GSO-NE of the present invention decreased somewhat at high temperatures, it was confirmed that only a slight decrease occurred even after 6 months at 4℃.

[0275] In summary, it can be confirmed that the GSO-NE of the present invention has excellent long-term durability and stability, as its durability does not change significantly under various storage conditions or periods at various temperatures.

[0276] In particular, as can be confirmed through FIG. 3E, the GSO-NE of the present invention is characterized by the optimized conditions of a particle size of 359.6 nm, a PDI of 0.24, and a zeta potential of -37.01 mV, and thus, it can be seen through these results that it is preferable to maintain a temperature lower than 4°C in order to maintain the stability or long-term durability of the GSO-NE of the present invention.

[0277]

[0278] Example 7. Confirmation of cytotoxicity and inflammatory activity in Vero E6 cells treated with GSO-NE.

[0279] The antiviral activity of the GSO-NE prepared in the above experimental example against the coronavirus HCoV-OC43 was confirmed. In particular, the safe and effective concentration of the GSO-NE of the present invention for treating HCoV-OC43 infection was confirmed.

[0280] Specifically, the above analysis was performed using MTT assay in Vero E6 cells infected with HCoV-OC43, which is shown in Fig. 3.

[0281] As a result, as can be confirmed in Fig. 3A, in terms of cytotoxicity, the GSO-NE of the present invention did not exhibit any toxic effect up to a treatment concentration of 5 μg / mL, and the cell viability was confirmed to be 80% even at a treatment concentration of 10 μg / mL, confirming that it is a safe concentration up to that concentration (Fig. 3A).

[0282] Also, as can be seen in Figure 3B, the IC of the GSO-NE of the present invention 50 The value was 18.08 μg / mL, and the ability of the GSO-NE of the present invention to respond to harmful viruses was confirmed by analysis of the morphology of Vero E6 cells after infection with HCoV-OC43 and treatment with GSO-NE in Fig. 3C. As can be seen from these results, replication of the virus occurred in cells infected with HCoV-OC43, causing damage to the host Vero E6 cells. However, when the GSO-NE of the present invention was treated, morphological changes in Vero E6 cells infected with HCoV-OC43, such as roundness, enlargement, swelling, and destruction of the cell layer, were confirmed to be reduced, thereby confirming the excellent antiviral activity of the GSO-NE of the present invention.

[0283] Furthermore, to further confirm the antiviral efficacy of GSO-NE, the mRNA level of the HCoV-OC43 N gene was determined in Vero E6 cells infected with HCoV-OC43. The nucleocapsid (N) protein is a structural component that binds to the genomic RNA while forming a helical capsid, and plays a very important role in the replication process of coronaviruses. Compared to the control group infected with HCoV-OC43, cells treated with the GSO-NE of the present invention showed a concentration-dependent decrease in the expression of the HCoV-OC43 gene at concentrations of 2.5 and 5 μg / mL, as shown in Fig. 3D.

[0284] Therefore, it can be seen that the GSO-NE of the present invention can effectively inhibit the replication of HCoV-OC43 in Vero E6 cells.

[0285]

[0286] Next, the effect of the GSO-NE of the present invention on inflammatory cytokines induced by HCoV-OC43 infection was confirmed.

[0287] Specifically, inflammatory cytokines such as interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and monocyte chemoattractant protein-1 (MCP-1) play essential roles in the body's defense mechanisms against viral infections, and are particularly associated with the cytokine storm observed in severe COVID-19 cases. These cytokines were identified with a focus on these cytokines because of their importance in inflammation, immune cell recruitment, and overall immune responses. Specifically, the mRNA expression levels of IL-6, TNF-α, IL-1β, and MCP-1 were quantified in Vero E6 cells by qRT-PCR, and the results are shown in Fig. 3E-H.

[0288] As a result, as can be confirmed in Figures 3E-H, it was confirmed that the mRNA expression level of related inflammatory cytokines significantly increased in cells infected with HCoV-OC43, and conversely, when GSO-NE of the present invention was treated together, it was confirmed that the RNA expression level of related cytokines significantly decreased in a concentration-dependent manner.

[0289] These results demonstrate that the GSO-NE of the present invention exhibits an effective inhibitory effect against HCoV-OC43 at a safe, non-cytotoxic concentration and has the effect of restoring cell morphology damaged by infection. In addition, GSO-NE suppresses the induction of inflammatory cytokines in Vero E6 cells infected with HCoV-OC43, indicating that GSO-NE has a therapeutic effect against HCoV-OC43 infection in Vero E6 cells.

[0290]

[0291] Example 8. Confirmation of the effect of GSO-NE on gene expression and pathways in Vero E6 cells.

[0292] To determine the effects of the present invention's GSO-NE on gene expression and pathways in Vero E6 cells, the effects of GSO-NE treatment on HCoV-OC43-infected Vero E6 cells were examined using RNA-Seq. The primer and antibody sequences used are shown in Tables 5 (SEQ ID NOs: 1 to 6) and 6, respectively.

[0293]

[0294]

[0295]

[0296]

[0297] As a result, as confirmed in Figures 4A-B, when analyzing the transcriptome, 146 genes were upregulated and 127 genes were downregulated, which could be confirmed using a volcano plot. In particular, in the case of ATP-binding cassette (ABC) transporters in HCoV-OC43 infection, a significant portion of related genes was confirmed to be downregulated, indicating inactivation of the pathway (Table 7 and Figure 4B).

[0298]

[0299]

[0300] These results suggest that treatment with the GSO-NE of the present invention can provide an innovative treatment by reversing gene expression within the ABC transporter pathway and reactivating the inactivated pathway.

[0301]

[0302] Example 9. Functional analysis of the ABCG gene in Vero E6 cells.

[0303] According to the results of Example 8 above, it was confirmed that treatment with the GSO-NE of the present invention reverses gene expression within the ABC transporter pathway and reactivates the inactivated pathway.

[0304] ABC transporters are a key membrane protein class that utilize the energy of ATP hydrolysis to actively transport various substrates across biological membranes. Through downstream RNA-seq analysis of DEGs, we focused on ABC target genes and identified subfamilies that play crucial roles in diverse physiological processes, including lipid metabolism, detoxification, drug resistance, and cellular homeostasis. Specifically, as shown in Figure 5A, we identified expression patterns of the ABC subfamily, including ABCA, ABCB, ABCC, ABCD, and ABCG.

[0305] As a result, the heatmap results showed that all subfamilies were significantly down-regulated in response to HCoV-OC43 infection, but treatment with the GSO-NE of the present invention effectively reversed these expression changes. In particular, ABCG subfamily genes were significantly down-regulated, indicating that the virus plays a role in regulating host gene expression or inducing immune evasion strategies.

[0306] Next, Figures 5B-C illustrate protein-protein interaction (PPI) analysis within the ABCG subfamily, including ABCA and ABCG transporters. Here, network visualization highlights interactions between different genes, including ABCG2, ABCG4, ABCG5, and ABCG8. This integrated analysis reveals a complex interplay between viral infection-induced changes in gene expression and the restorative effects of GSO-NE on ABC transporters, particularly the ABCG subfamily.

[0307]

[0308] Example 10. Reversal of ABCG inhibition by GSO-Ne in Vero E6 cells

[0309] The results of Examples 8 and 9 above show that the antiviral activity of the GSO-NE of the present invention is closely related to ABCG, a subfamily of the ABC transporter superfamily.

[0310] Accordingly, in order to confirm the effect of HCoV-OC43 infection on the regulation of ABCG transporter expression, the mRNA expression amount of Vero E6 cells treated with the GSO-NE of the present invention was quantified using qRT-PCR, and the results are shown in Figs. 6A-E (Figs. 6A-E).

[0311] As a result, it was confirmed that the gene expression levels of ABCG1, ABCG2, ABCG4, ABCG5, and ABCG8 were significantly reduced after infection with HCoV-OC43 in Vero E6 cells. In contrast, it was confirmed that the expression levels of the five genes increased after treatment with the GSO-NE of the present invention.

[0312] Next, looking at the results of protein expression analysis performed through immunoblotting (Fig. 6F), it was possible to reconfirm that the expression of ABCG1, ABCG5, and ABCG8 proteins was restored in the above results, and in particular, the expression of ABCG1 was confirmed to be significantly higher than that of the control group, ABCG5, and ABCG8.

[0313] These results demonstrate the important role of ABCG transporters in coronavirus infections such as HCoV-OC43, and in particular, suggest that the GSO-NE of the present invention can be utilized as an excellent antiviral agent against coronaviruses such as HCoV-OC43 by reversing and reactivating the expression of a subfamily of ABCG transporters, particularly a subfamily such as ABCG1.

[0314]

[0315] In summary, GSO-NE, which was optimized as a nanoemulsion using the GSO of the present invention, has excellent antiviral activity against coronaviruses such as HCoV-OC43 at a non-cytotoxic concentration in Vero E6 cells. In particular, transcriptome analysis results showed that the gene expression of the ABC transporter G subfamily was significantly downregulated in Vero E6 cells infected with HCoV-OC43 after treatment with GSO-NE, confirming that the GSO-NE of the present invention can be used as an antiviral therapeutic agent by reversing and reactivating the expression of the ABC transporter G subfamily. These results suggest a potential mechanism by which the GSO-NE of the present invention exerts its antiviral effect and demonstrate that it also affects cellular transport processes that are important for viral infection. In addition, the advantages of using nanoemulsions in drug delivery, such as improved stability, improved bioavailability, and targeted administration, suggest that GSO-NE has potential as a potential therapeutic agent for HCoV-OC43.

[0316]

[0317] From the above description, those skilled in the art will understand that the present invention can be implemented in other specific forms without altering its technical concept or essential characteristics. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be interpreted as encompassing all changes or modifications derived from the meaning and scope of the following claims and their equivalent concepts, rather than the detailed description above.

Claims

1. ABCG transporter activator containing ginseng seed oil.

2. In the first paragraph, the ginseng seed oil is an ABCG transporter activator characterized by inhibiting the expression of the ABCG transporter.

3. In the first paragraph, the ABCG transporter activator is at least one selected from the group consisting of ABCG1, ABCG2, ABCG4, ABCG5, and ABCG8.

4. In the third paragraph, the ABCG transporter activator is at least one selected from the group consisting of ABCG1, ABCG5, and ABCG8.

5. An ABCG transporter activator according to claim 1, wherein the ginseng seed oil is in the form of an oil-in-water nanoemulsion formulation.

6. An ABCG transporter activator according to claim 1, wherein the ginseng seed oil-in-water nanoemulsion further comprises a surfactant.

7. An ABCG transporter activator according to claim 1, wherein the surfactant is an amphiphilic surfactant, a hydrophobic surfactant, or a combination thereof.

8. An ABCG transporter activator according to claim 7, wherein the amphipathic surfactant comprises Tween 80.

9. An ABCG transporter activator according to claim 7, wherein the hydrophobic surfactant comprises Span 80.

10. An ABCG transporter activator according to claim 7, wherein the amphipathic surfactant and the hydrophobic surfactant are mixed with ginseng seed oil in a ratio of 1:1 to 1:

4.

11. An ABCG transporter activator according to claim 10, wherein the amphipathic surfactant and the hydrophobic surfactant are mixed with ginseng seed oil in a ratio of 1:

2.

12. An ABCG transporter activator according to claim 1, characterized in that the ginseng seed oil-in-water nanoemulsion is prepared at an ultrasonic treatment time of 15 to 25 minutes and an ultrasonic treatment amplitude of 50 to 70%.

13. In the first paragraph, the ginseng seed oil-in-water nanoemulsion is characterized by a reduced droplet size and an improved zeta potential compared to nanoemulsions formulated under other conditions, thereby significantly improving colloidal stability, which is an ABCG transporter activator.

14. An antiviral composition against a coronavirus comprising an ABCG transporter activator according to any one of claims 1 to 13.

15. An antiviral composition for coronavirus in claim 14, wherein the coronavirus is at least one selected from the group consisting of SARS-CoV, SARS-CoV-2, MERS-CoV, and human coronavirus-OC43 (HCoV-OC43).

16. An antiviral composition for coronavirus, characterized in that the composition has antiviral activity by increasing the expression of the ABCG transporter suppressed by the coronavirus and reactivating the inactivated pathway in accordance with claim 14.

17. An antiviral composition for coronavirus, characterized in that the composition further has anti-inflammatory activity in accordance with claim 14.

18. An antiviral composition for coronavirus, wherein the antiviral composition further comprises silinidin in the 14th paragraph.

19. A pharmaceutical composition for preventing or treating a coronavirus infectious disease, comprising an ABCG transporter activator according to any one of claims 1 to 13.

20. A food composition for preventing or improving a coronavirus infectious disease, comprising an ABCG transporter activator according to any one of claims 1 to 13.

21. A feed composition for preventing or improving coronavirus infectious disease, comprising an ABCG transporter activator according to any one of claims 1 to 13.

22. A method for producing an antiviral composition against coronavirus comprising an ABCG transporter activator comprising a ginseng seed oil oil-in-water nanoemulsion, the method comprising a step of formulating ginseng seed oil into an oil-in-water nanoemulsion.

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